Dihydroschizozygin and process for producing the same



United StatesPatent O 3,251,831 DHIYDROSCHIZOZYGIN AND PROCESS FOR PRODUCING THE SAME Ulrich Renner, Riehen, near Basel, Switzerland, assignor t Geigy Chemical Corporation, Greenburgh, N.Y., a corporation of Delaware No Drawing. Filed June 12, 1 9,63, Ser. No. 287,223 Claims priority, application Switzerland, June 14, 1962, 7,175/62; Feb. 13, 1963, 1,774/ 63 7 Claims. (Cl. 260-293.4)

This invention relates to dihydroschizozygin and to a process for producing the same.

It has been found that a good yield of an alkaloid mixture can be extracted from parts, particularly from the roots and bark, of Schizozygia cafiaeoides (Boj.) Baill., which belongs to the Apocynaceaefamily and grows in East Africa. This alkaloid mixture consists chiefly of a previously unknown, uniform alkaloid that can be crystallized, which will now be termed schizozygin and which has valuable therapeutical properties. Its antipyretic and bronchospasmolytic action are particularly marked; that it also has an action on the central nervous system can be seen from its anaesthesia and analgesics potentiating effect.

As extracting agents there can be used either polar solvents such as lower alkanols with at most three carbon atoms, if necessary with addition of an alkanoic acid with at most three carbon atoms, or non-polar or weakly polar solvents such as chloroform, methylene chloride or ben zene, respectively, or ether-type solvents such as diethyl ether, dibutyl ether, dioxan or tetrahydrofuran, in the presence of water and, if necessary, in the presence of a Weak inorganic or organic base.

On using polar solvents, the schizozygin is extracted partially as such and particularly as salt of the acids Which are present in the plant material; and on addition of low alkanoic acids as salt of the latter. The presence of Water and, optionally, of an inorganic base such as sodium bicarbonate, sodium acetate, sodium or potassium carbonate, calcium hydroxide or ammonia or an organic base which boils below 100, e.g. trimethyl amineor triethyl amine, i.e. the moistening of the plant material with water or with aqueous solutions of suitable bases before extraction with at most Weak polar solvents brings about the hydrolysis of the salts of schizozygin, which are present in the plant, or liberates schizozygin from its salts respectively and thus permits its extraction of the free base.

The extraction of schizozygin can be performed e.g. according to the process which is described in detail in the following.

Ground and dried parts, particularly the roots and bark of the trunk of Schizozygia caflaeoides, are extracted with an alkanol containing 1-3 carbon atoms, optionally with the addition of an alkanoic acid containing 1-3 carbon atoms, such as acetic acid in amounts up to 1% by volume, the extract is concentrated under reduced pressure, diluted with water, optionally with the addition of an alkanoic acid containing 1-3 carbon atoms, and the alkanol is completely distilled off under reduced pressure. An acid to neutral aqueous phase containing chlorine ions, in particular concentrated hydrochloric acid or an aqueous solution of an alkali or alkaline earth metal chloride is added to the acid aqueous solution of the alkaloid mixture which remains, preferably after clarification of the latter by filtration over a little kieselguhr, the crude schizozygin hydrochloride which precipitates is separated and the base is liberated, e.g. by treatment of the hydrochloride with ammonia or with an aqueous solution of an alkali or earth alkali hydroxide. The schizozygin obtained can be purified, e.g. by recrystallization from methanol; if desired, the crude hydrochloride can also be even previously purified, e.g. by crystallization from a low alkanol.

Instead of precipitating the schizozygin as hydrochloride from the acid aqueous solution of the alkaloid mixture, it can also be precipitated as such from this solution. This is done preferably by first clarifying this solution by filtration over a little kieselguhr, and then bringing the pH of the solution to 8-10, e.g. by the addition of ammonia or an aqueous solution of an alkali or earth alkali hydroxide or carbonate. The precipitated mixture of alkaloids is isolated, e.g. by filtration or centrifugation and the schizozygin is obtained therefrom by crystallization from an alkanol containing 1-3 carbon atoms.

In order to extract the schizozygin by a different mode of operation, "the ground parts of the plant of Schizozygia cafiaeoides are first moistened with a diluted aqueous sodium carbonate solution, i.e. the material can be sprayed and mixed Well. .The plant material treated in this way is extracted e.g. with chloroform, the chloroform extract is concentrated and then methanol is added which causes the schizozygin to crystallize out. The schizozygin obtained by the process according to the invention crystallizes from chloroform/methanol in colorless needles which melt at 192-194; +155 (c.=1, chloroform).

Elementary analysis shows that it has the formula 20 20 3 2 Calculated: C, 71.42; H,5.99; N, 8.33. Found: C,

Group analyses for OCH NCH and CCH are negative, the Labat test for the presence of the grouping 208 m (log e 4.38), 269 m (log 6 3.99) and 313 mu (log 5 3.97). In the short wave range of the infra-red spectrum (in KBr) there is a strong band at 6.05 1. C O lactam).

schizozygin hydrochloride can be obtained either direct according to the process of the invention or by the addition of hydrochloric acid to an acetic acid or alcoholic solution of schizozygin. From methanol it crystallises in needles which melt at over 250 (under carbonisation). Other salts with inorganic or organic acids can be obtained in an analogous manner. Schizozygin perchlorate crystallises from methanol in colourless needles which melt at 252-255 (with decomposition). I

On reacting schizozygin with methyl iodide, a methoiodide is obtained which crystallises as colourless leaflets from methanol which decompose at over 280 under carbonisation.

Catalytic hydrogenation of the methochloride produced from the above methoiodide (Emde-degradation), with the take-up of about double the molar amount of hydrogen, produces a tertiary base and, on oxidising this with chromic acid, butyric acid among others is obtained.

In the presence of platinum catalyst, schizozygin absorbs a substantially equimolar amount of hydrogen and loses a double bond. A tetrahydro derivative is formed as side product which, in contrast to schizozygin and dihydro schizozygin, contains an NH group and a CH group bound to a carbonatom.

Finally, a ditertiary base is obtained on reduction of schizozygin by means of lithium aluminium hydride. In Z-methoxyethanol, a PK value of 4.29 was found for schizozygin.

From the spectra, group analyses, the Labat test and the above degradation reactions, the following Formula I results for schizozygin:

As expected, Hof-mann degradation of the methoiodide mentioned above with potassium tert. butylate, produced a tertiary amine with an additional dOLlblCybOIld. This shows a proton resonance spectrum, the signs of which confirm the above Formula I because they can be completely correlated to a 3-vinyl indoline derivative, derived from Formula II below, in which formula the v-values (s=singlet, d=doublet) attributed to the various protons are given:

If the 3-vinyl indoline derivative of Formula II is treated with potassium tert. butylate, then an isomer is obtained which, already in the Hofmann degradation reaction, is also found as by-product and which, moreover, is characterised by an extended chromophoric system with UV maxima at 230, 298, 333 and 346 m corresponding to a double bond conjugated with the benzene nucleus. It contains a methyl group appearing in the resonance spectrum with a doublet at 7' 7 .95, which can also be traced according to Kuhn-Roth. The vinyl radical in the 3- position of the indole nucleus can thus be rearranged ,to storm the 3-ethylidene group which increases the probability of the'accuracy of Formulae I and II. In this connection, it is pointed out that in the Ho'fmann degradation of the methoiodide, the assumption that the unsaturated 6-membered ring containing an allylamine grouping is split, can be excluded because the Emde degradation process which, in this case, is possible only if there is an allylamine group, can be performed with theproduct of the Hofmann degradation process, i.e. identical products are obtained if, on the one hand, the 3-vinyl indoline derivative, of Formula II, is again quaternised and then hydrogenated (degradation according to Emde and hydrogenation of the vinyl group) and, on the other hand, the degradation product of scrizozygin according to Emde is quaternised, degraded according to Hofmann and the product obtained is hydrogenated.

Furthermore, a compound which is distinguished from the compound of Formula I by a much greater centrally active component in the form of an unexpectedly higher anaesthesia potentiating action and lack of bronchospasmolytic action can be obtained if schizozygin of the general Formula I given above is catalytically hydrogenated until substantially an equimolar amount of hydrogen is taken up and the compound of formula I of the formula IV in practice comprises the stepsof (a) Extracting bark of Schizozygia cafiaeoz'des with an alcohol selected from the group consisting of methanol and ethanol,

b) Acidifying the resulting alcoholic extract with a dilute aqueous to concentrated acetic acid of from about 5 to strength and separating the liquid from the resulting solid phase,

(c) Adjusting the pH of the resulting acidified liquid phase to about 10,

(d) Separating the resulting precipitate from the mother liquor,

(e) Dissolving the precipitate in a hot alcohol selected from the group consisting of methanol and ethanol,

(f) Cooling the resulting alcoholicsolution, whereby a product having a melting point of 192 C. crystallizes, and separating the latter fromthe mother liquor,

(g) Dissolving the said crystalline product in glacial acetic acid and hydrogenating the dissolved product with hydrogen in the presence of a platinum oxide hydrogenation catalyst at room temperature, and

(h) Recovering the resulting hydrogenation product of Formula III from the hydrogenation mixture.

As mentioned hereinbefore, either stem bark or root bark is used as starting material in step (a).

Methanol is the, preferred solvent in steps (a) and (e). In step (c) the pH is adjusted to 10 by adding to the said acidified mixture a base selected from the'group consisting of ammonia, alkali metal hydroxide, alkali metal carbonate and alkali metal hydrogen carbonate.

Both schizozygin and dihydr'o-schizozygin form monoacidic salts with inorganic and organic acids, such as bydrochloric acid, hydrobrornic acid, sulfuric acid, phosphoric acid, methane sulfonic acid, acetic acid, lactic acid, oxalic acid, succinic acid, fumaric acid, maleic acid, malic acid, tartaric acid, citric acid, benzoic acid, salicyclic acid, phenyl acetic acid and with mandelic acid.

The following examples further illustrate the processes according to the invention but are by no means the only The hydrogenation is performed, for example, in

methods of performing the same. The parts are given as parts by weight; their relationship to parts by volume is as that of grams to cubic centimeters. The temperatures are given in degrees centigrade. Percentages are by weight.

Example 1 7000 parts of ground bark from the trunk of Schizozgia cafiaeoides are exhaustively extracted with 30,000 parts by volume of absolute methanol, the extract is concentrated in vacuo to about 2,000 parts by volume, about 3,000 parts by volume of aqueous acetic acid are added and the remaining methanol is distilled ott in vacuo. The solution which remains is clarified by filtration over kieselguhr, brought to a pH of about 10 with ammonia and the precipitated mixture of the crude product (153 parts) is filtered off under suction. This is washed with water and taken up in hot methanol. 77 parts of schizozygin crystallize, M.P. 190-192". To produce the hydrochloride, 1 part of schizozygin is dissolved in 20 parts of 2 N acetic acid and, While cooling and stirring, 2 parts by volume of concentrated hydrochloric acid are added to the solution. The precipitated hydrochloride is filtered off and crystallised from methanol. 1 part of schizozygin hydrochloride is obtained. It decomposes above 250 with carbonisation.

Example 2 500 parts of finely ground bark from the roots of Schizozygia cafiaeoides are extracted three times while stirring with 2,000 parts by volume of methanol each time. I The methanolic extract is concentrated in vacuo to 100 parts by volume and then diluted with aqueous 1 N 'acetic acid to 100 parts by volume. The acid solution is Example 3 500 parts of finely ground bark from the roots of Schizozygia cafiaeoides are extracted three times while stirring with 2,000 parts by volume of absolute methanol each time. The methanolic extract is concentrated to 100 parts by volume in vacuo and taken up in 300 parts by volume of aqueous 2 N acetic acid. The acid solution is filtered over a little kieselguhr and 50 parts by volume of concentrated hydrochloric acid are added. After 24 hours, about 7 parts of crude schizozygin hydrochloride separate as a dark red resinous product. This filtered oil, dissolved in water, ammonia is added to the solution until the reaction is alkaline, the precipitated bases are removed by filtration and the schizozygin is obtained therefrom by crystallisation from methanol.

Example 4 '500 parts of the ground bark from the trunk of Schizozygia cafiaeoides are percolated with 3,000 parts by volume of methanol which contains parts by volume of acetic acid. After concentrating the extract in vacuo to about 100 parts by volume, it is diluted with 1,000 parts by volume of water, the solution obtained is clarified by filtration over kieselguhr, the pH is brought to about 10 with 2 N sodium hydroxide solution and the crude alkaloids are filtered oflf under suction. These are washed with water and crystallised from methanol. 5 parts of schizozygin are obtained. M.P. 191193.

Example 5 500 parts of ground bark from the roots of Schizozygia caflaeoides are sprayed with a 1% solution of sodium carbonate and mixed well. The drug material prepared in this way is then percolated with 5,000 parts by volume of chloroform. The chloroform extract is concentrated of schizozygin crystallise out.

Example 6 4 parts of schizozygin are dissloved in parts by' volume of glacial acetic acid and the solution is hydrogenated in the presence of 0.5 part of platinum oxide according to Adams at 25 and a pressure of 746 Torr until no more hydrogen is taken up. (Hydrogen absorp tion: 314 parts by volume.) After filtering off the catalyst and washing with a little Water, the filtrate is diluted with 200 parts by volume of water, made alkaline wtih ammonia and extracted three times with 100 parts by volume of ether each time. The ether extracts are combined, dried with sodium sulphate, evaporated to dryness and the residue is crystallised from ether. 3.2 parts of dihydro-schizozygin are obtained. M.P. 190491", [a] +29.4 (in chloroform; c.=1).

For the production of the tartrate 1 part of dihydroschizozygin is dissolved in 10 parts of methylene chloride and an equivalent amount of 1 N alcoholic tartaric acid is added. Then the solution is concentrated to dryness.

Example 7 3.36 parts of schizozygin are dissolved in 250 parts by volume of dioxan and hydrogenated in the presence of 2.5 parts of Raney nickel at 20 and normal pressure (744 mm. Hg). After ca. 21 hours no further hydrogen is taken up. The catalyst is filtered oil, the filtrate is concentrated to dryness and the residue is crystallised from ether. 2.5 parts of dihydro-schizozygin are obtained, M.P. l89191.

I claim:

7 1. The compound of the formula with a pharmaceutically acceptable acid.

3. A process for the production of the compound of the Formula III comprising (a) extracting bark of Schizozygia cafiaeoides with an alcohol selected from the group consisting of methanol and ethanol,

about 5 to 100% strength, and separating the liquid from the resulting solid-phase, (c) adjusting the pH of the resulting acidified liquid phase to aboutlO,

(d) separating the resulting precipitate from the mother liquor, (e) dissolving the precipitate in a hot alcohol selected from the group consisting of methanol and ethanol, (f) cooling the resulting alcoholic solution, whereby a product having a melting point of 190192 crystallises, and separating the latter from the mother liquor, (g) dissolving the said crystalline product in glacial acetic acid and hydrogenating the dissolved product with hydrogen in the presence of a platimum oxide hydrogenation catalyst at room temperature, and

8 (h) recovering the'resulting hydrogenation product of Formula III from the hydrogenation mixture. 4. Aprocess as described in claim 3 wherein the bark used in step (a) is ground stem bark.

5. A process as described in claim 3 wherein the bark used in step (a) is ground root bark.

6. A process asdescribed in claim 3 wherein the pH i is adjusted in step (c) by adding to the said acidified mixture a base selected from the group consisting of ammonia, alkali metal hydroxide, alkali metal hydrogen HENRY R. JILES, Acting PrimaryExaminer 

1. THE COMPOUND OF THE FORMULA 